Antibacterial activity in three Chaetoceros microalgae species cultures by using antibiotics

Diatoms, such as Chaetoceros, grow in a mutualistic relationship with bacteria. However, in some cases, it is necessary to grow them in bacteria-free cultures. To reduce bacterial load, antibiotics are used, and on certain occasions it is necessary to use a mixture with more than one antibiotic. This work aimed to obtain a quick and effective protocol to reduce the bacterial load and evaluate the response of three Chaetoceros species with aquacultural importance. Single and mix antibiotics were used. Microalgal and bacterial growth was measured. The growth parameters for diatoms showed that the significantly highest cell concentration was for C. muelleri (3.15 x106 cells mL-1) and the lowest values to C. calcitrans (2.98 x106 cells mL-1). The significantly highest growth rate was for C. calcitrans (0.77 divisions per day), and the lowest values for Chaetoceros sp. (0.60 divisions per day). The growth parameters for heterotrophic bacteria showed that the significantly highest bacterial load was for Chaetoceros sp. (19.16 x106 CFU (Colony-Forming Units) mL-1) and the lowest values were for C. calcitrans (12.23 x106 CFU mL-1). The growth rate of the heterotrophic bacteria present in Chaetoceros cultures was similar among the three studied species. Streptomycin® and sulfate G41® produced a partial reduction of bacterial load. The most effective treatment for all three species was the use of an antibiotic mix composed of ampicillin® (250 μg mL-1), kanamycin® (200 μg mL-1), neomycin® (50 μg mL-1), and streptomycin® (100 μg mL-1) for three days. The mix prepared with the highest antibiotic concentration produced a reduction of bacteria (100%) for three days; however, it also induced a significant reduction of the growth of the three Chaetoceros species

Decreasing of bacterial content in Isochrysis galbana cultures by using some antibiotics

The axenic microalgae cultures are a difficult task and they are hard to maintain. Microalgae cultures with reduced bacterial load can be an option to axenic microalgae cultures to produce compounds with biotechnological and pharmaceutical potential. Also they can be used for cryopreservation and in biochemical, physiology, ecology and genetic studies. The aim of this study was to develop a protocol to decrease the bacterial load in an Isochrysis galbana culture, through washes by centrifugation and the administration of various antibiotics (ampicillin, neomycin, kanamycin, chloramphenicol, sulphate G418, streptomycin, and carbencillin), at several doses and combinations. The concentrations of heterotrophic bacteria and I. galbana cell densities were monitored daily. Maximum non-lethal concentration and lethal concentration 50% (LC50) were calculated. Individually, antibiotics and washes by centrifugation failed to reduce bacterial load, but their combination removed bacteria from the cultures. Peak survival (84.6 ± 1.4%) and reduction of bacterial load in I. galbana cultures were effected with the combination of 5 washes by centrifugation and administration of a cocktail, comprising ampicillin, kanamycin, neomycin, and streptomycin at 48 h. Values of maximum non-lethal concentration varied from 75 to 106 µg mL-1 and LC50 between 194 and 332 µg mL-1, thus, our protocol is an effective and rapid method of producing I. galbana cultures with reduced bacterial load.La obtención de cultivos axénicos es una tarea laboriosa y resultan difíciles de mantener. Los cultivos de microalgas con carga bacteriana reducida pueden ser una alternativa a los cultivos axénicos de microalgas para producir compuestos con potencial biotecnológico y farmaceútico. También éstos pueden ser usados para la criopreservación, y en estudios bioquímicos, fisiológicos, ecológicos y genéticos. El propósito de este estudio fue desarrollar un protocolo para reducir la carga bacteriana en cultivos de Isochrysis galbana por medio de lavados por centrifugación y la administración de varios antibióticos (ampicilina, neomicina, kanamicina, cloranfenicol, sulfato G418, estreptomicina y carbencilina), adicionados en varias dosis y combinaciones. Todos los tratamientos fueron realizados por triplicado. La concentración de bacterias heterotróficas y la densidad de células de I. galbana fue evaluada diariamente. Se calculó la concentración no letal máxima y la concentración letal (LC50). De forma individual, los antibióticos y los lavados por centrifugación no fueron efectivos para reducir la carga bacteriana, pero su combinación removió las bacterias de los cultivos. El máximo de sobrevivencia (84,6 ± 1,4%) y la reducción de la carga bacteriana en I. galbana fue efectiva con la combinación de 5 lavados por centrifugación y la administración de un coctel de ampicilina, kanamicina, neomicina, y estreptomicina por 48 h. Los valores de concentración no letal máxima varían entre 75 a 106 µg mL-1 y LC50 se encontró entre 194 y 332 µg mL-1, por lo anterior, este protocolo resulta ser un método efectivo y rápido para obtener cultivos de I. galbana con carga bacteriana baja